SBIR Phase I: Next-Generation Capillary Electrophoresis with Mass Spectrometry for Biopharmaceutical and Biomedical Applications

这个小型企业创新研究(SBIR)项目的更广泛影响/商业潜力将是开发一种强大的毛细管电泳仪，具有用于生物医学研究、生物制药开发、临床诊断和系统生物学的质谱仪检测功能。毛细管电泳法是一种功能强大的液体分离技术，在生物分析中具有广阔的应用前景。它是完成人类基因组计划的主要工具，在一定程度上导致了DNA测序市场。同时具有光学和质谱学检测的CE系统将使人们能够更深入地了解控制生化过程的分子动力学，如蛋白质表达、放松调控以及与其他生化物种的相互作用，这可能导致在药物开发、早期诊断和治疗致命人类疾病(如癌症、糖尿病和神经退行性疾病)方面取得突破。拟议的技术将通过提高产品表征的效率来提高质量改进，从而为生物制药药物开发公司带来好处。这个SBIR第一阶段项目的智力优势是开发了用于蛋白质表征的下一代毛细管电泳电喷雾电离质谱仪(CE-ESI-MS)。CE-ESI-MS可以对包括蛋白质在内的多种生化物种进行高效分离和表征。然而，由于缺乏强大、易用的仪器，利用CE-ESI-MS的固有优势一直很困难，这使得常规应用中采用CE-ESI-MS的情况并不常见。本研究将开发和评估一种用于蛋白质表征的新型CE-ESI-MS仪器。拟议的文书将采用原创性创新，以克服现有技术的局限性并超越其能力。将开发一种用于样品和缓冲液操作的新型流体纳米管，以允许从1微升或更少的样品体积进行分析。纳米港将采用全自动化设计，操作员干预较少。这将有助于样本有限的应用，例如在临床分析中。此外，该仪器将利用混合电喷雾电离接口同时进行光学和MS检测，这是现有技术所不能提供的。将毛细管电泳法的超高效率与光学检测和质谱学的分子鉴定相结合，将成为生物分析的有力工具。这一组合将代表着相对于最先进技术的重大进步，并增强了询问生物样品中蛋白质及其生化功能的手段。拟议的创新将显著增强CE-ESI-MS在快速、高效和深入生物分析方面的稳健性和能力。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be the development of a robust capillary electrophoresis instrument with mass spectrometry detection for biomedical research, biopharmaceutical development, clinical diagnostics, and system biology. Capillary electrophoresis (CE) is a powerful liquid separation technology with remarkable potentials for bioanalysis. It was the workhorse tool for the completion of human genome project that, in part, led to the DNA sequencing market. A CE system with both optical and mass spectrometry detection will enable a deeper understanding of molecular dynamics governing biochemical processes, such as protein expression, deregulation, and interaction with other biochemical species, which could lead to breakthroughs in drug development, early diagnosis, and treatments for deadly human diseases such as cancer, diabetics, and neurodegenerative disorders. The proposed technology will offer benefits to biopharmaceutical drug development companies by increasing the efficiency of product characterization for quality improvement. It also will allow efficient routine proteomics analysis and other bioanalytical applications.The intellectual merit of this SBIR Phase I project is to develop a next-generation capillary electrophoresis with electrospray ionization mass spectrometry (CE-ESI-MS) for protein characterization. CE-ESI-MS allows high efficiency separation and characterization of several biochemical species including proteins. However, leveraging the intrinsic advantages of CE-ESI-MS has been difficult due to lack of robust, easy-to-use instrumentation, which has made adoption for routine applications uncommon. This research will develop and evaluate a novel CE-ESI-MS instrument for protein characterization. The proposed instrument will employ original innovations to overcome the limitations of the incumbent technologies and exceed their capabilities. A novel fluidic nanoport for sample and buffer manipulation will be developed to allow analysis from 1 microL or less sample volume. The nanoport will employ a fully automated design with less operator intervention. This will be useful for sample-limited applications such as in clinical analysis. In addition, the instrument will utilize a hybrid electrospray ionization interface for simultaneous optical and MS detection, which no existing technology offers. Combining CE's ultrahigh efficiency with robust quantitative characteristics of optical detection and the molecular identification of mass spectrometry will be a powerful tool for bioanalysis. This combination will represent a significant advancement over the state-of-the-art and enhance the means of interrogating proteins and their biochemical functions in biological samples. The proposed innovations will significantly enhance the robustness and capabilities of CE-ESI-MS for fast, efficient, and deep bioanalysis.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.